Fig. 1. Genetic constructs and expression analysis of cyanobacterial FBP/SBPase in C. reinhardtii.
(a) Schematic representation of expression cassettes used to generate FBP/SBPase expressing lines both with or without mVenus (YFP) fused at the C-terminus. To drive gene expression the hybrid promoter HSP70-RBCS2 was used, while the FBP/SBPase was targeted into chloroplast by exploiting the PsaD target peptide sequence (Photosystem I Subunit D). Both expression cassettes contained the Aph VIII gene confer resistance to the antibiotic paromomycin. Both FBP/SBPase proteins either fused or not to YFP carried a C-terminal S-tag for immunodetection and protein purification. (b) Immunoblot against S-tag showing the FBP/SBPase accumulation in transformed lines without YFP as reporter protein. (c) Fluorescence screening of transformed lines with FBP/SBPase_YFP. The best five lines were subjected to immunoblot against YFP confirming the presence of the protein are reported in the Panel c insert.
